Abstract
The Gibson Assembly is a popular method for molecular cloning which has been developed specifically to join several fragments together in a specific order, without the constraint of restriction enzyme sites. This method is based on the assembly of overlapping fragments, generally produced by PCR, and then combining them using three enzymes: a 5′ exonuclease, a DNA polymerase, and a DNA ligase, in an isothermal reaction. Here, we describe this method, including the design of primers for the generation of the overlapping fragments and the assembly; to this end, we provide an example involving joining two fragments in a single plasmid.
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Avilan, L. (2023). Assembling Multiple Fragments: The Gibson Assembly. In: Scarlett, G. (eds) DNA Manipulation and Analysis. Methods in Molecular Biology, vol 2633. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3004-4_4
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DOI: https://doi.org/10.1007/978-1-0716-3004-4_4
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-3004-4
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